Transfected HeLa cells were fixed with 2% paraformaldehyde at room temperature for 10 min at 24 h after transfection and permeabilized with 1% TritonX-100 at 4°C for 15 min. Fixed cells were incubated with 2% BSA at 37°C for 30 min for blocking. Anti-V5 antibody (Thermo Fisher Scientific, 1:500) was added at 4°C overnight. Alexa 546 conjugated-anti-mouse IgG antibody (Thermo Fisher Scientific, 1:600) and 1 μg/ml of Hoechst 33342 (Sigma-Aldrich) were added at room temperature for 45 min. Fluorescence was observed with a fluorescent microscope BZ-9000 BIOREVO (KEYENCE, Osaka, Japan).
Alexa 546 conjugated anti mouse igg antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
Alexa Fluor 546 conjugated anti-mouse IgG antibody is a secondary antibody used in immunodetection applications. It is designed to bind to primary mouse antibodies, allowing for fluorescent detection and visualization of target antigens.
Automatically generated - may contain errors
Lab products found in correlation
Anti v5 antibody, by Thermo Fisher Scientific (1 mentions)
Hoechst 33342, by Merck Group (1 mentions)
Mouse anti n cadherin antibody, by BD (1 mentions)
Zsm710, by Zeiss (1 mentions)
Af6000, by Leica (1 mentions)
Fv1000, by Olympus (1 mentions)
Ab51451, by Abcam (1 mentions)
Anti zeb1, by Cell Signaling Technology (1 mentions)
Anti gapdh 6c5, by Merck Group (1 mentions)
Tetramethylrhodamine isothiocyanate tritc conjugated phalloidin, by Merck Group (1 mentions)
Anti jarid2, by Novus Biologicals (1 mentions)
Anti zeb2, by Active Motif (1 mentions)
Anti e cadherin, by Thermo Fisher Scientific (1 mentions)
Anti fibronectin, by Merck Group (1 mentions)
Anti e cadherin, by BD (1 mentions)
Anti vimentin, by Abcam (1 mentions)
4 protocols using alexa 546 conjugated anti mouse igg antibody
1
Immunofluorescence of Transfected HeLa Cells
2
Immunofluorescence analysis of FAK and N-cadherin
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Cells were fixed in 4% paraformaldehyde and blocked in 5% skim milk/phosphate buffer saline with 0.1% Tween-20 (PBST). After washing in PBST, cell samples were incubated with rabbit anti-FAK(p397) antibody (Abcam, Cambridge, UK) or mouse anti-N-cadherin antibody (BD Biosciences, San Jose, CA, USA) in 5% skim milk/PBST, washed with PBST twice, and incubated with Akexa488-conjugated anti-rabbit IgG antibody or Alexa546-conjugated anti-mouse IgG antibody (Thermo Fisher Scientific, Waltham, MA, USA). Cell specimens were also stained with rhodamine phalloidin (Cytoskelton, Inc., Denver, CO, USA). Mounted cell specimens were analyzed with a confocal microscope (ZSM710; Carl Zeiss, Jena, Germany).
3
Indirect Immunofluorescence Analysis of HBs Antibody
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Indirect immunofluorescence analysis was performed as previously described47 (link). The anti-HBs antibody was used as the primary antibody at 1:200 dilution, and the secondary antibody, Alexa 546-conjugated anti-mouse IgG antibody (Invitrogen, California, USA), was used at 1∶1000 dilution. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). Fluorescence imaging was performed with AF6000, a fluorescence microscope (Leica, Wetzlar, Germany), or FV1000, a laser scanning confocal microscope (Olympus, Tokyo, Japan).
4
Immunoblotting and Fluorescence Microscopy Assay
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Immunoblotting was performed as described previously [7] (link). Anti-JARID2 (#NB100-2214, Novus Biologicals), anti-E-cadherin (#610181, BD Transduction Lab), anti-Fibronectin (SAB4500974, Sigma), anti-Vimentin (ab8069, Abcam), anti-ZEB1 (#3396, Cell Signaling), anti-ZEB2 (#61096, Active Motif), anti-phosphorylated SMAD3 (ab51451, Abcam) and anti-GAPDH (6C5, Millipore) antibodies were used. To detect the morphological changes of the cells, A549 or HT29 cells were stained with 0.4% crystal violet (Waldeck). To allow direct fluorescence of actin cytoskeleton, the cells were stained with 0.25 µM tetramethylrhodamine isothiocyanate (TRITC)-conjugated phalloidin (Sigma). For indirect immunofluorescence, the specimens were incubated with anti-E-cadherin antibody and treated with Alexa546-conjugated anti-mouse IgG antibody (Invitrogen). Nuclei were visualized with 4′,6-diamidino-2-phenylindole (DAPI).
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